目的 构建人p53RFP基因启动子序列不同截短片段的荧光素酶报告基因载体,研究启动子的转录活性.方法 PCR扩增人p53RFP基因启动子区域不同长度的目的片段,克隆到pGL3-Basic中,构建启动子区域3个不同截短片段的荧光素酶报告基因载体,经双酶切和基因测序鉴定正确后,转染HEK293细胞,双荧光素酶报告基因检测系统检测荧光素酶活性.结果 成功构建了p53RFP启动子不同截短片段的荧光素酶报告基因载体,经双荧光素酶报告基因检测分析,3个启动子片段均具有转录活性.结论 成功构建了人p53RFP基因启动子荧光素酶报告基因载体,为研究p53RFP基因的转录调控机制提供了实验基础.%Objective To construct the luciferase reporter gene vector containing human p 53RFP gene promoter, and study the transcriptional activity of the promoter .Methods The different fragments of p53RFP promoter were gener-ated by PCR, and subsequently cloned into pGL 3-Basic to construct luciferase reporter vectors of p 53RFP gene promot-er.The resulting vectors were verified by double enzyme digestion and DNA sequencing .The luciferase activities were de-tected by dual luciferase reporter system in HEK 293 cells.Results Three luciferase reporter gene vectors containing p53RFP gene promoter were successfully constructed .The dual luciferase reporter assay showed that all the three vectors had promoter activity.Conclusion The luciferase reporter gene vectors containing p 53RFP promoter region are success-fully constructed, which provides a basis for study on transcriptional regulatory mechanisms of p 53RFP gene.
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